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Fig. 4. Reduced Rab5 and enhanced Rab7 content of MBSA-MDP-treated cells is
sufficient to drive lysosomal targeting. To determine the role of Rab5 and
Rab7 content of MBSA-MDP-treated cells in lysosomal targeting of
Salmonella, reconstitution of the phagosome-lysosomes transport assay
was carried out in permeabilized cells in the presence of cytosol prepared
from MBSA-MDP-treated cells or untreated cells containing in vitro prenylated
Rab proteins as described in the Materials and Methods. Cells were incubated
for 60 minutes at 37°C to allow transport. Subsequently, formation of the
bacteria biotin-avidin-HRP complex was measured to determine the transport of
the Salmonella to lysosomes. Each point represents the
mean±s.d. from three independent experiments, and these are expressed
as relative units of transport to lysosomes.
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